title: Functional integration of grafted neural stem cell-derived dopaminergic neurons monitored by optogenetics in an in vitro Parkinson model.
abstract: Intrastriatal grafts of stem cell-derived dopamine (DA) neurons induce behavioral recovery in animal models of Parkinson's disease (PD), but how they functionally integrate in host neural circuitries is poorly understood. Here, Wnt5a-overexpressing neural stem cells derived from embryonic ventral mesencephalon of tyrosine hydroxylase-GFP transgenic mice were expanded as neurospheres and transplanted into organotypic cultures of wild type mouse striatum. Differentiated GFP-labeled DA neurons in the grafts exhibited mature neuronal properties, including spontaneous firing of action potentials, presence of post-synaptic currents, and functional expression of DA D₂ autoreceptors. These properties resembled those recorded from identical cells in acute slices of intrastriatal grafts in the 6-hydroxy-DA-induced mouse PD model and from DA neurons in intact substantia nigra. Optogenetic activation or inhibition of grafted cells and host neurons using channelrhodopsin-2 (ChR2) and halorhodopsin (NpHR), respectively, revealed complex, bi-directional synaptic interactions between grafted cells and host neurons and extensive synaptic connectivity within the graft. Our data demonstrate for the first time using optogenetics that ectopically grafted stem cell-derived DA neurons become functionally integrated in the DA-denervated striatum. Further optogenetic dissection of the synaptic wiring between grafted and host neurons will be crucial to clarify the cellular and synaptic mechanisms underlying behavioral recovery as well as adverse effects following stem cell-based DA cell replacement strategies in PD.
authors.affiliation: Experimental Epilepsy Group, Wallenberg Neuroscience Center, Lund University Hospital, Lund, Sweden.
authors.forename: Jan
authors.initials: J
authors.lastname: Tønnesen
authors.forename: Clare L
authors.initials: CL
authors.lastname: Parish
authors.forename: Andreas T
authors.initials: AT
authors.lastname: Sørensen
authors.forename: Angelica
authors.initials: A
authors.lastname: Andersson
authors.forename: Cecilia
authors.initials: C
authors.lastname: Lundberg
authors.forename: Karl
authors.initials: K
authors.lastname: Deisseroth
authors.forename: Ernest
authors.initials: E
authors.lastname: Arenas
authors.forename: Olle
authors.initials: O
authors.lastname: Lindvall
authors.forename: Merab
authors.initials: M
authors.lastname: Kokaia
mesh_terms: D000200:Action Potentials / Q000502:physiology; D000818:Animals; D048430:Cell Shape; D000075402:Channelrhodopsins; D004298:Dopamine / Q000378:metabolism*; D066298:In Vitro Techniques; D008636:Mesencephalon / Q000473:pathology / Q000503:physiopathology; D051379:Mice; D008954:Models, Biological; D058953:Neural Stem Cells / Q000166:cytology / Q000637:transplantation*; D009474:Neurons / Q000166:cytology / Q000637:transplantation*; D055095:Optics and Photonics / Q000379:methods*; D010300:Parkinson Disease / Q000503:physiopathology / Q000628:therapy*; D013569:Synapses / Q000378:metabolism
chemical_list: D000075402:Channelrhodopsins; D004298:Dopamine